Tyramide Signal Amplification (TSA), also known as Catalyzed Reporter Deposition (CARD), offers an effective way to efficiently enhance signal and detection capabilities for low-abundance targets in immunocytochemistry (ICC), immunohistochemistry (IHC), and in situ hybridization (ISH) applications.
Kits |
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Cat. No. | Product Name / Activity |
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7523 | TSA Vivid™ Fluorophore Kit 520 |
Signal amplification kit for use in ISH, ICC and IHC | |
7526 | TSA Vivid™ Fluorophore Kit 570 |
Signal amplification kit for use in ISH, ICC and IHC | |
7527 | TSA Vivid™ Fluorophore Kit 650 |
Signal amplification kit for use in ISH, ICC and IHC | |
Other |
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Cat. No. | Product Name / Activity |
6241 | Biotinyl Tyramide |
Reagent widely used for signal amplification in IHC and FISH | |
6457 | Cyanine 3 Tyramide |
Orange fluorescent reagent widely used for signal amplification in IHC and FISH | |
6458 | Cyanine 5 Tyramide |
Red fluorescent reagent widely used for signal amplification in IHC and FISH | |
7236 | Digoxigenin Tyramide |
Reagent used for Tyramide Signal Amplification in IHC, ICC and ISH | |
6456 | Fluorescein Tyramide |
Green fluorescent reagent widely used for signal amplification in IHC and FISH |
Tyramide Signal Amplification (TSA), also known as catalyzed reporter deposition (CARD), offers an effective way to efficiently boost your signal and detection capabilities for low-abundance targets in IHC, ICC and FISH. HRP catalyzes the conversion of labeled inactive tyramide into a reactive radical, which then binds to adjacent tyrosine residues greatly enhancing the density of labeling. Tyramide reagents can be conjugated with a dye for fluorescent detection, or a hapten for antibody-based probing.
TSA can be used to enhance the signal of multiple targets on one sample by deactivating HRP using a peroxidase quenching buffer, then carrying out another round of TSA using a different set of antibodies from a different host species. Alternatively, the antibodies for the initial round of TSA can be stripped, leaving the covalently bound tyramides. TSA can then be repeated for the desired number of targets.
Figure 1: Tyramide Signal Amplification Principles: A primary and secondary antibody are used to label a tissue or cell sample. The secondary antibody is pre-conjugated to horseradish peroxidase (HRP), which in the presence of H2O2, catalyzes a labeled tyramide substrate into a highly reactive species that covalently bind to tyrosine residues on the proteins in close proximity to the antibodies and HRP, thus providing signal amplification.
Our TSA Vivid Fluorophore Kits are engineered for increased brightness and improved performance in ICC, IHC and FISH applications. They are specifically designed for optimal performance in the RNAscope™ Multiplex Fluorescent v2 Assay, enabling visualization of gene expression at the single cell level.
Key features of TSA Vivid Fluorophore Kits:
Figures 2 and 3 demonstrate the improved brightness of TSA Vivid dyes in the RNAscope™ Multiplex Fluorescent v2 Assay versus a leading competitor.
Figure 2: 3-plex RNAscope™ Multiplex Fluorescent v2 Assay on HeLa cells with the leading competitor dyes (left) and the corresponding TSA Vivid dyes 520, 570 and 650 (right). All dyes were used at 1:1500 dilution. Markers shown are Polr2a in green, PPIB in red and UBC in white. Nuclei are counter-stained with DAPI (Cat. No. 5748).
Figure 3: 3-plex RNAscope™ Multiplex Fluorescent v2 Assay on human lung cancer tissue with the leading competitor dyes (left) and the corresponding TSA Vivid dyes 520, 570 and 650 (right). All dyes were used at 1:1500 dilution. Cancer immuno-oncology markers were probed: IDO-1 in green, KRT-19 in red and PD-1 in white. Nuclei are counter-stained with DAPI (Cat. No. 5748).
Find out how our customers have used and what they think of our 5-star TSA product Biotinyl Tyramide (Cat. No. 6241), as they share experimental details and images of their results.
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